Influence of Cytokeratin 18 on subcellular distribution of LRP16 protein

Objective:To construct recombinant expression vector of LRP16 carryied GFP gene and study the influence of Cytokeratin 18(KRT18)on subcellular distribution of LRP16 fusion protein,Western blot was initially used to further accredit the influence.Methods:The primarily structural sequence of LRP16 was recombined in pEGFP-N1 vector,inserting in the N-termini of GFP(green fluorescent protein),and then transferred or cotransferred with KRT18 into MCF-7,NIH3T3 cell line,fluorescent signal was detected using fluorescent microscope.Overexpressing KRT18 and downregulation of endogenous KRT18 expression by RNA interference in MCF-7 cell and Hela cell lines,Western blot was used to investigate effects of down-regulation on subcellular distribution of LRP16.Results:Successfully construct the recombinant expression vector of pEGFP-LRP16.The green fluorescene was inspired from cellular nucleolus in MCF-7 cells.LRP16-GFP was located mainly in cytoplasm in NIH3T3 cells.KRT18 protein could mediate LRP16 to distribute from cellular nucleus to cytoplasm.Western blot obtained the same results.Conclusion:Keratin18 is one of the most important proteins that interacts with LRP16 and play an important fuction in subcellular distribution of LRP16.These results provide essential clues for further investigation of the functional regulation of LRP16.